Use of propionibacterium granulosum in the treatment of necrotizing enterocolitis

Abstract

The application provides application of Propionibacterium granulosum in treatment of necrotizing enterocolitis, and belongs to the technical field of microorganisms. The application finds for the first time that the Propionibacterium granulosum can be used for treating necrotizing enterocolitis. The application proves by a mouse experiment that the Propionibacterium granulosum significantly improves the survival rate of mice with necrotizing enterocolitis, and effectively reduces intestinal pathological damage of the necrotizing enterocolitis. The Propionibacterium granulosum can reduce intestinal epithelial barrier damage, promote intestinal epithelial barrier repair and inhibit intestinal tissue inflammatory damage. Mechanism research proves that the treatment effect depends on the biological activity of live bacteria, and has bacteria specificity - the same genus of Propionibacterium acnes does not have the curative effect. The application provides a brand-new strategy for treatment of necrotizing enterocolitis.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology, specifically relating to the application of Propionibacterium granulosum in the treatment of necrotizing enterocolitis. Background Technology

[0002] Necrotizing enterocolitis (NEC) is the most common acute intestinal disease in the neonatal period, with high morbidity and mortality rates in severely ill infants, and is a major cause of death in premature infants. Currently, there is a lack of specific treatment strategies for NEC targeting its etiology. Early treatment of NEC is mainly supportive, and surgical intervention is necessary when intestinal perforation or conservative medical treatment fails. Some surviving infants suffer from long-term sequelae such as intestinal stenosis, short bowel syndrome, and neurodevelopmental delay, severely impacting their quality of life. Therefore, the prevention and treatment of NEC remains a very urgent task.

[0003] The pathogenesis of neonatal endocarditis (NEC) remains unclear, but recent mainstream views suggest that multiple factors contribute to its development, including immature intestinal development, impaired intestinal barrier function, gut microbiota dysbiosis, and excessive immune responses. In the neonatal gut, early colonization of gut microbiota is crucial for maintaining intestinal barrier integrity and regulating intestinal immune function. Studies have shown that gut microbiota dysbiosis in preterm infants often precedes the onset of clinical symptoms and is a key initiating factor in NEC pathogenesis. Therefore, correcting gut microbiota dysbiosis, repairing the intestinal barrier, and suppressing intestinal inflammatory responses through exogenous supplementation with specific probiotics has become an important direction in NEC treatment. Thus, screening for specific bacterial strains with definite gut-protective effects has significant clinical translational and scientific value for NEC treatment. Summary of the Invention

[0004] In view of this, in order to provide a new treatment strategy for NEC, this invention has demonstrated that Propionibacterium granulosum has good therapeutic effects in improving the survival rate of NEC mice, improving intestinal pathological damage, enhancing intestinal epithelial barrier function, and inhibiting intestinal immune inflammation.

[0005] The first aspect of the present invention provides particulate Propionibacterium ( Cutibacterium granulosum Its application in the preparation of drugs for treating NEC.

[0006] In some implementations, the NEC is an NEC that occurs in newborns.

[0007] In some implementations, the drug is a drug that reduces intestinal pathological damage caused by NEC.

[0008] In some implementations, the drug for reducing NEC intestinal pathological damage is a drug that reduces intestinal edema, reduces intestinal wall gas accumulation, and reduces the degree of damage to small intestinal villi.

[0009] In some implementations, the drug is a drug that enhances the intestinal epithelial barrier function and reduces the level of inflammatory factors in intestinal tissue.

[0010] In some implementations, the enhancement of intestinal epithelial barrier function includes reducing intestinal epithelial barrier damage and promoting intestinal epithelial barrier repair.

[0011] In some implementations, the inflammatory factors include one or both of TNF-α and IL-6.

[0012] In some embodiments, the dosage form of the drug includes, but is not limited to, solutions, lyophilized powders, capsules, tablets, granules, drops, pills, suppositories, powders, suspensions, or emulsions.

[0013] In some embodiments, the drug also contains excipients and / or carriers.

[0014] In some embodiments, the excipients and / or carriers include, but are not limited to, any one or a combination of at least two of the following: fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, colorants, pH regulators, antioxidants, antibacterial agents, or buffers.

[0015] In some implementations, a freeze-drying protectant may be added to the freeze-dried powder.

[0016] In some implementations, the capsule includes a soft capsule and a hard capsule.

[0017] In some embodiments, the tablets include, but are not limited to, excipients.

[0018] In some implementations, the granules may contain, but are not limited to, starch, sucrose, and maltodextrin.

[0019] In some embodiments, oil may be selected as the solvent in the drops. Selectable oils include, but are not limited to, food oils.

[0020] In some embodiments, the particulate Propionibacterium in the drug is a live bacterium.

[0021] A second aspect of the present invention provides a pharmaceutical composition comprising particulate Propionibacterium acnes and its pharmaceutically acceptable carrier and / or excipients.

[0022] In some embodiments, the viable count of Propionibacterium granules in the pharmaceutical composition is a therapeutically effective amount.

[0023] In some implementations, the effective therapeutic dose is 1×10⁻⁶. 7 CFU-1×10 9 CFU.

[0024] In some implementations, the effective therapeutic dose is 1×10⁻⁶. 7 CFU.

[0025] A third aspect of the present invention provides a method for treating NEC, the method comprising administering the pharmaceutical composition described in the second aspect of the present invention.

[0026] The advantages and beneficial effects of this invention are as follows: This invention provides the application of *Propionibacterium granulosum* in the treatment of NEC. This invention is the first to discover that *Propionibacterium granulosum* can be used to treat NEC. This invention comprehensively evaluates the therapeutic effect of *Propionibacterium granulosum* based on mouse body weight, survival rate, gross intestinal pathology, and NEC severity score. This invention elucidates for the first time the role and mechanism of *Propionibacterium granulosum* in enhancing the intestinal epithelial barrier function and inhibiting inflammatory damage in NEC, providing a novel strategy for the treatment of NEC. Attached Figure Description

[0027] Figure 1 Schematic diagram of NEC modeling in newborn mice under different bacterial treatments Figure 2 This was used to assess the severity of NEC in each group of mice. Figure 2 A is a statistical graph showing the body weight of mice in each group. Figure 2 B represents the survival curves of mice in each group. Figure 2 C shows gross pathological images of the intestines of mice in each group. Figure 2 Image D shows HE staining of the terminal ileum of mice in each group. Figure 2 E is a statistical graph of HE pathological disease scores for each group of mice.

[0028] Figure 3 This was to assess the intestinal epithelial barrier function of each group. Figure 3 A represents the detection of intestinal permeability in each group of mice. Figure 3 B shows immunofluorescence images of the tight junction proteins Claudin1, Occludin, ZO-1, and adhesion junction protein E-cadherin in the intestinal epithelial cells of each group of mice.

[0029] Figure 4 This section describes the apoptosis and proliferation of intestinal epithelial cells in each group. Figure 4 Image A shows representative images of TUNEL immunofluorescence staining of the terminal ileum of mice in each group. Figure 4 B represents the quantitative statistical count of TUNEL-positive cells per intestinal villus in each group of mice. Figure 4C shows the immunofluorescence images of cleaned Caspase 3 (CC3) in the terminal ileum of mice in each group. Figure 4 D represents the quantitative statistical count of CC3-positive cells per intestinal villus in each group of mice. Figure 4 E shows immunofluorescence images of the PCNA in the terminal ileum of mice in each group. Figure 4 F represents the quantitative statistical percentage of PCNA-positive cells in each crypt unit of mice in each group. Figure 4 G represents the quantitative statistics of the height of the terminal ileum villi and the depth of the crypts in each group of mice.

[0030] Figure 5 The study aimed to detect the TLR4 / NF-κB signaling pathway and intestinal inflammatory factors in the intestinal tissue of mice in each group. Figure 5 Image A shows immunofluorescence images of TLR4 in the intestinal tissue of mice in each group. Figure 5 B shows immunofluorescence images of P-NF-κB p65 in the intestinal tissue of mice in each group. Figure 5 CD is a statistical graph showing the levels of TNF-α and IL-6 in the intestinal tissue of mice in each group.

[0031] Figure 6 for C. granulosum Safety assessment in healthy mice. Figure 6 A represents the levels of feces from mice in each group. C. granulosum A statistical chart of relative abundance. Figure 6 B is a statistical graph showing the body weight of mice in each group. Figure 6 C represents the survival curves of mice in each group. Figure 6 D shows gross pathological images of the intestines of mice in each group. Figure 6 E shows HE staining images of the terminal ileum of mice in each group. Figure 6 F is a statistical graph of the HE pathological disease scores of mice in each group. Figure 6 G represents the intestinal permeability test of mice in each group. Detailed Implementation

[0032] The present invention will be further described below with reference to embodiments. The following description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any other way. Any person skilled in the art may make equivalent modifications to the disclosed technical content to create equivalent embodiments. Any simple modifications or equivalent changes made to the following embodiments based on the technical essence of the present invention without departing from the scope of the invention are all within the protection scope of the present invention.

[0033] Example The culture of the particulate Propionibacterium was 1×10⁻⁶. 8The concentration was found to be CFU / ml. The specific method involved taking frozen Propionibacterium granules, adding them to 10 ml of minced meat carbohydrate broth, and placing them in an anaerobic bag in a 37°C incubator. After 48 hours of incubation, the Propionibacterium granules were serially diluted 10-fold to 10... -9 100 μL of bacterial culture from each gradient was spread onto Columbia blood agar plates and anaerobic incubated for 48 hours. The bacterial colony count was then observed, and the fixed OD was calculated. 600 The number of CFUs at a given value was used to fit the growth curve of *Propionibacterium granulosum*. Activated and well-grown *Propionibacterium granulosum* were then analyzed using the measured OD value. 600 A fixed amount of bacterial culture was taken, and the bacterial cells were collected by centrifugation at a concentration of 1×10⁻⁶. 7 The CFU cells were resuspended in PBS and administered by gavage. The culture can be added to breast milk or formula as an additive, or prepared as a probiotic preparation as a lyophilized powder.

[0034] Formula and preparation method of minced meat carbohydrate broth culture medium: 30.0g casein peptone, 10.0g beef extract, 5.0g yeast extract, 5.0g dipotassium hydrogen phosphate, 0.5g L-cysteine ​​hydrochloride, 0.0005g resazurin, 4.0g glucose, 1.0g cellobiose, 1.0g maltose, 1.0g starch, pH 7.0±0.2 (25℃). Weigh 57.7g of the above ingredients into 1L of distilled water, heat to boiling and continue boiling for more than 1 minute. Add minced beef granules to the culture medium to 1 / 3 of the liquid height. Autoclave at 121℃ for 20 minutes. Cool to below 50℃ and add 5μg / ml heme chloride and 50μg / ml vitamin K1.

[0035] Propionibacterium acnes was anaerobically cultured in modified PYG liquid medium for 48 hours. Growth curves of Propionibacterium acnes were plotted using the method described above, and the OD values ​​were measured. 600 A fixed amount of bacterial culture was taken, and the bacterial cells were collected by centrifugation at a concentration of 1×10⁻⁶. 7 The CFU cells were resuspended in PBS and administered by gavage.

[0036] Modified PYG liquid culture medium formulation and preparation method: 5.0g tryptone, 5.0g peptone, 10.0g yeast extract, 5.0g beef extract, 5.0g glucose, 2.0g dipotassium hydrogen phosphate, 1.0ml Tween 80, 0.01g CaCl2·2H2O, 0.02g MgSO4·7H2O, 0.4g sodium bicarbonate, 0.08g sodium chloride, 1mg resazurin, pH 7.2±0.2 (25℃). Weigh 33.6g of the above ingredients into 1L of distilled water, heat to boiling and continue boiling for at least 1 minute, autoclave at 121℃ for 20 minutes, cool to below 50℃, and add 5μg / ml heme chloride, 1μg / ml vitamin K1, and 0.5mg / ml cysteine ​​hydrochloride.

[0037] 1. Test Methods 1.1 Test strains Propionibacterium granulosum ( Cutibacterium granulosum CCUG 67158 was purchased from Wuhan Gray Algae Biotechnology Co., Ltd., product number HZB902753. Propionibacterium acnes (… Cutibacteriumacnes ATCC 6919 was purchased from Wuhan Gray Algae Biotechnology Co., Ltd., product number HZB358661.

[0038] 1.2 Experimental Animals C57BL / 6J newborn mice were provided by Jilin Qianhe Model Biotechnology Co., Ltd.

[0039] 1.3 Experimental Grouping There are a total of 5 groups: control group, NEC model group (NEC group), NEC model combined group, and NEC model combined group. Cutibacterium granulosum Processing group (NEC+) C. granulosum (Group), NEC model combined with thermal inactivation Cutibacterium granulosum Processing group (NEC+HI- C. granulosum (Group) and NEC model combined Cutibacterium acnes Processing group (NEC+) C.acnes Group).

[0040] 1.4 Test Methods Control group newborn mice were breastfed after birth without any special treatment. The NEC mouse model was induced for 4 days using a "hypoxia-formula-gavage feces from surgically induced NEC puppies" method. Newborn mice on day 7 (P7) were selected for modeling. After the modeling began, they were separated from their mothers and gavaged with formula milk five times daily at a ratio of 2:1 (Abbott Similac infant formula and PetAg Esbilac puppy milk powder). Each ml of formula milk contained 12.5 μl of fecal homogenate from surgically induced NEC puppies. During the modeling period, newborn mice were subjected to hypoxia twice daily, placed in a hypoxic chamber of 5% O2-95% N2 for 10 minutes, followed by cold stimulation at 4 ℃ for 10 minutes. NEC+ C. granulosum The mice in group P5 were given daily gavage starting with NEC induction. C. granulosum (1×10) 7 CFU (50 μl) was administered once, until NEC modeling was completed. NEC+HI- C. granulosum In addition to inducing NEC, mice in the group were administered HI- by gavage daily. C. granulosum (PBS resuspension) C. granulosum After heat inactivation treatment at 65℃ for 2 hours, 1×10 7 CFU (50 μl) once. NEC+ C.acnes In mice group P5, after induction of NEC, daily gavage was initiated. C.acnes (1×10) 7 CFU (50 μl) was administered once daily until the NEC model was completed. The body weight and survival of mice in each group were monitored daily. Terminal ileum tissue from newborn mice was collected at P11, fixed with paraformaldehyde, and used for HE staining and immunofluorescence detection, or cryopreserved for the detection of inflammatory factor levels. The modeling procedure is as follows: Figure 1 As shown.

[0041] 1.5 Detection Indicators 1.5.1 Mouse weight monitoring, mortality statistics, and gross intestinal pathology 1.5.2 HE pathological disease scoring criteria for intestinal tissue: 0 points, no damage; 1 point, damage to the tips of the villi or mild separation of the submucosa and lamina propria; 2 points, rupture of the villi in the middle and / or moderate separation of the submucosa and lamina propria; 3 points, complete rupture of villi and / or severe separation and / or edema of the submucosa; 4 points, transmural damage.

[0042] 1.5.3 Mouse intestinal permeability: Mice were fasted the night before sampling. The next day, they were given 10 mg / ml 70-kDa FITC-glucan (44 mg / 100g body weight) by gavage. Four hours later, blood was collected by cardiac puncture. The serum was separated and diluted with an equal volume of PBS. The fluorescence intensity of FITC-glucan in the serum was measured by a multi-mode microplate reader. The excitation wavelength was set to 493 nm and the emission wavelength to 517 nm.

[0043] 1.5.4 Expression of intercellular junctions in intestinal epithelial cells: After fixing the terminal ileum tissue of mice in each group, paraffin sections were prepared, and immunofluorescence staining was used to detect the expression of intestinal epithelial cell tight junction proteins Claudin1, Occludin, ZO-1, and adhesion junction protein E-cadherin.

[0044] 1.5.5 Detection of intestinal epithelial cell apoptosis and expression of apoptosis-related protein CC3: TUNEL immunofluorescence staining was used to detect apoptosis in intestinal epithelial cells of mice in each group. Cell nuclei were counterstained with DAPI, and images were captured using a fluorescence microscope. TUNEL-positive cells were quantified using a blinded method, expressed as the number of TUNEL-positive cells per intestinal villus. Immunofluorescence staining was used to detect the expression of CC3, an apoptosis-related protein in intestinal epithelial cells, and the number of CC3-positive cells per intestinal villus was quantitatively counted.

[0045] 1.5.6 Detection of intestinal epithelial cell proliferation: Immunofluorescence staining was used to detect the expression of PCNA, a protein associated with the proliferation of intestinal epithelial cells, and the percentage of PCNA-positive cells in each crypt unit was quantitatively analyzed.

[0046] 1.5.7 Detection of the TLR4 / NF-κB signaling pathway in intestinal tissue: Immunofluorescence staining was used to detect the expression of TLR4 and P-NF-κB p65 in intestinal tissue.

[0047] 1.5.8 Levels of intestinal inflammatory factors TNF-α and IL-6: Terminal ileum tissue from each group was homogenized with sterile PBS, and the supernatant was collected by centrifugation. The levels of TNF-α and IL-6 in the intestinal tissue of each group were detected by ELISA.

[0048] 1.5.9 Feces C. granulosum Abundance detection: Fecal samples were collected from the colon during sampling. Fecal genomic DNA was obtained from each group using a fecal genomic DNA extraction kit, then diluted to a DNA concentration of 1 ng / μl, and analyzed using SYBR and... C. granulosumqPCR detection was performed using specific primers, and expression levels were normalized relative to bacterial 16S v3-v4. 2 -ΔΔCT Method C. granulosum Quantitative analysis of abundance.

[0049] 1.6 Statistical Methods Experimental data were statistically analyzed using GraphPad Prism 9 software. Quantitative data were expressed as mean ± standard deviation. Comparisons between two groups of means were performed using... t For comparisons among multiple groups, one-way ANOVA was used. P < 0.05 indicates a statistically significant difference.

[0050] 2. Experimental Results 2.1 Changes in mouse body weight: The results are as follows Figure 2 As shown in Figure A, the body weight of mice in the control group showed a steady increase with age, while the body weight gain of mice in the NEC group slowed significantly. Compared with the NEC group, the NEC+ group showed a significantly slower increase in body weight. C. granulosum On day 4 of modeling, the body weight of mice in the NEC+HI- group increased significantly. C. granulosum On day 4 of modeling, the body weight of mice in the group was lower than that of NEC+ mice. C. granulosum The group showed a significant decrease, NEC+ C.acnes On day 4 of modeling, the body weight of the mice in the control group was not significantly different from that in the NEC group.

[0051] 2.2 Mouse survival status: The results are as follows Figure 2 As shown in B. After modeling, the survival rate of mice in the control group was 100%, the survival rate of mice in the NEC group was 60%, and the survival rate of mice in the NEC+ group was 60%. C. granulosum The survival rate of mice in the NEC+HI- group was 90%. C. granulosum The survival rate of mice in the group was 70%, NEC+ C.acnes The survival rate of mice in the group was 60%, suggesting that... C. granulosum Live bacteria can improve the survival rate of NEC mice.

[0052] 2.3 Gross pathological findings of the intestines of mice in each group are as follows: Figure 2 As shown in C, the HE staining and pathological disease score statistics of the terminal ileum tissue of mice in each group are as follows. Figure 2 As shown in D and 2E, C. granulosum Treatment reduced NEC intestinal pathological damage. Gross intestinal pathology showed reduced intestinal edema and significantly reduced intestinal wall gas accumulation. HE pathology indicated reduced damage to small intestinal villi, with the intestinal mucosa and lamina propria remaining relatively intact. Furthermore, the NEC severity score was significantly reduced, while the HI- C. granulosum and C.acnes Treatment with this method did not significantly improve the intestinal pathology of NEC mice. These results demonstrate that... C. granulosumLive bacteria can significantly reduce the severity of NEC.

[0053] 2.4 Detection of intestinal epithelial barrier function: The results are as follows Figure 3 A and Figure 3 As shown in B, compared to the NEC group, NEC+ C. granulosum The intestinal permeability of the mice in the group was significantly reduced, and the expression of intestinal epithelial cell tight junction proteins Claudin1, Occludin1, ZO-1 and adhesion junction protein E-cadherin was significantly increased, while the expression of NEC+HI- C. granulosum Group and NEC+ C.acnes No significant changes were observed in the group, suggesting... C. granulosum The treatment enhanced the intestinal epithelial barrier function in NEC mice.

[0054] 2.5 Detection of intestinal epithelial cell apoptosis and intestinal epithelial cell proliferation: The results of TUNEL staining and CC3 immunofluorescence are as follows: Figure 4 AB and Figure 4 As shown in CD, C. granulosum Treatment significantly reduced the number of TUNEL-positive and CC3-positive cells in the intestinal tissue of NEC mice, suggesting that this strain can inhibit apoptosis of intestinal epithelial cells in NEC mice. PCNA immunofluorescence results are as follows: Figure 4 As shown in EF, C. granulosum Treatment significantly increased the proportion of PCNA-positive cells, suggesting that this strain can promote the proliferation of intestinal epithelial cells in NEC mice. Meanwhile, C. granulosum Treatment restored the height of the small intestinal villi and the depth of the crypts in NEC mice. Figure 4 G), reflecting the structural changes in the intestinal epithelium after the restoration of the apoptosis / proliferation balance. These changes are observed in NEC+HI- C. granulosum Group and NEC+ C.acnes No similar effect was observed in the control group of mice. Therefore, C. granulosum The treatment effectively promotes the repair of the NEC intestinal epithelial barrier.

[0055] 2.6 Detection of TLR4 / NF-κB signaling pathway and intestinal inflammatory factors in mouse intestinal tissue: The immunofluorescence results of TLR4 are as follows: Figure 5 As shown in A, compared to the NEC group, NEC+ C. granulosum TLR4 expression was significantly decreased in the group. Immunofluorescence results for p-p65 are as follows: Figure 5 As shown in B, compared to the NEC group, NEC+ C. granulosum The p-p65 signal in the nuclei of intestinal epithelial cells of this group was significantly reduced, indicating that C. granulosum Treatment with this agent inhibits the phosphorylation and nuclear translocation of NF-κB p65. These results indicate that... C. granulosum Live bacteria have a significant inhibitory effect on the overactivation of the TLR4 / NF-κB signaling axis.

[0056] Results of mouse intestinal tissue inflammatory factor levels as follows Figure 5 As shown in CD, compared to the NEC group, NEC+ C. granulosum The group significantly reduced the levels of intestinal inflammatory factors TNF-α and IL-6, suggesting C. granulosum The treatment effectively inhibited NEC-induced intestinal inflammation.

[0057] 2.7 C. granulosum Safety assessment in healthy mice: qPCR detection control group and C. granulosum Group (starting from P7, once daily gavage) C. granulosum (4 days in total, no other special treatment) Mouse feces C. granulosum The abundance, the results are as follows Figure 6 As shown in A, gavage C. granulosum The strain subsequently colonized the mouse gut.

[0058] healthy mice by gavage C. granulosum It does not affect its weight gain and survival rate, such as Figure 6 BC; both gross intestinal pathology and HE pathology scores indicated no pathological changes in the intestine, such as Figure 6 DF; also does not increase intestinal permeability in mice, such as Figure 6 As shown in G. Therefore, C. granulosum This strain has good safety profile as it neither affects the general condition of healthy mice nor causes intestinal damage.

[0059] The description of the above embodiments is only applicable to understanding the method and core ideas of the present invention. It should be noted that those skilled in the art can make various improvements and modifications to the present invention without departing from its principles, and these improvements and modifications will also fall within the protection scope of the claims of the present invention.

Claims

1. Application of Propionibacterium granulosum in the preparation of drugs for treating necrotizing enterocolitis.

2. The application according to claim 1, characterized in that, The drug is used to reduce intestinal pathological damage caused by necrotizing enterocolitis.

3. The application according to claim 2, characterized in that, The drugs mentioned that reduce intestinal pathological damage in necrotizing enterocolitis are those that reduce intestinal edema, reduce intestinal wall gas accumulation, and reduce the degree of damage to small intestinal villi.

4. The application according to claim 1, characterized in that, The drug is used to reduce intestinal epithelial barrier damage in necrotizing enterocolitis, promote intestinal epithelial barrier repair, and reduce the level of inflammatory factors in intestinal tissue.

5. The application according to claim 4, characterized in that, The drugs that reduce intestinal epithelial barrier damage in necrotizing enterocolitis are those that reduce intestinal permeability and restore the expression of intercellular junction proteins in intestinal epithelial cells.

6. The application according to claim 4, characterized in that, The drugs that promote the repair of the intestinal epithelial barrier are those that reduce intestinal epithelial cell apoptosis and promote intestinal epithelial cell proliferation.

7. The application according to claim 4, characterized in that, The inflammatory factors include one or both of TNF-α and IL-6.

8. The application according to claim 1, characterized in that, The dosage forms of the drug include solutions, lyophilized powders, capsules, tablets, granules, or drops.

9. The application according to claim 1, characterized in that, The drug also contains excipients and / or carriers.

10. The application according to claim 1, characterized in that, The Propionibacterium granules in the drug are live bacteria.

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